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Leishmaniasis is a neglected and widespread parasitic disease that can lead to serious health problems. The conventional method in diagnostic health clinics is direct smear preparation of the lesion and staining with standard Giemsa to visualize the amastigote stage and by culturing the organism in an NNN (Novy-MacNeal-Nicolle) to observe the promastigote form of the parasite. In the case of urban-type leishmaniasis, microscopic diagnosis is sometimes not possible due to the reduction of amastigotes in patients' wounds. Because most endemic areas are located in regions that do not have access to laboratories equipped with molecular tools, access to a rapid test to diagnose the disease is essential. In this study, for the first time for DNA extraction, the scalpel used for sampling was washed and extracted by boiling method. Also, the LAMP technique in this study was modified so that the test can be performed in 10 min and the results can be recognized by color. We used four microscopic methods, conventional PCR, real-time PCR, and LAMP, to diagnose urban-type leishmaniasis and compared the results of these methods with each other. The sensitivity and specificity of LAMP were higher than other techniques used. Therefore, it allows rapid diagnosis for timely treatment of the disease to control the primary reservoir host more quickly in ACL as humans are the principal source of infection. This test is performed at a high-speed and is cost-effective. For its convenience, this test is highly recommended to be used in endemic areas.
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Leishmania , Leishmaniose Cutânea , Humanos , Leishmania/genética , Leishmaniose Cutânea/parasitologia , Técnicas de Diagnóstico Molecular , Técnicas de Amplificação de Ácido Nucleico , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Pentavalent antimonials (Sb(V)) such as meglumine antimoniate (Glucantime®) and sodium stibogluconate (Pentostam®) are used as first-line treatments for leishmaniasis, either alone or in combination with second-line drugs such as amphotericin B (Amp B), miltefosine (MIL), methotrexate (MTX), or cryotherapy. Therapeutic aspects of these drugs are now challenged because of clinical resistance worldwide. METHODS: We reviewedthe recent original studies were assessed by searching in electronic databases such as Scopus, Pubmed, Embase, and Web of Science. RESULTS: Studies on molecular biomarkers involved in drug resistance are essential for monitoring the disease. We reviewed genes and mechanisms of resistance to leishmaniasis, and the geographical distribution of these biomarkers in each country has also been thoroughly investigated. CONCLUSION: Due to the emergence of resistant genes mainly in anthroponotic Leishmania species such as L. donovani and L. tropica, as the causative agents of ACL and AVL, respectively, selection of an appropriate treatment modality is essential. Physicians should be aware of the presence of such resistance for the selection of proper treatment modalities in endemic countries.
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Antiprotozoários , Leishmaniose Cutânea , Leishmaniose , Gluconato de Antimônio e Sódio/uso terapêutico , Antiprotozoários/farmacologia , Antiprotozoários/uso terapêutico , Biomarcadores , Resistência a Medicamentos/genética , Humanos , Leishmaniose/tratamento farmacológico , Leishmaniose Cutânea/tratamento farmacológico , Antimoniato de Meglumina/uso terapêuticoRESUMO
AIM: Vulvovaginal candidiasis (VVC), is a common fungal infection that remains a global concern. The objectives of this study were molecular identification and assessment of the antifungal susceptibility profile of Candida species, causing VVC in southeast Iran. METHODS: A cross-sectional investigation was carried out on 119 nonpregnant females suspected of VVC between February 2019 and May 2021. Yeast samples were characterized to the species level by conventional and molecular methods. All Candida isolates were examined for in vitro susceptibility profile to six conventional antifungal drugs using Clinical and Laboratory Standards Institute guidelines. RESULTS: Out of 119 subjects, 52 (43.7%) cases were affected by VVC, out of whom 11 (21.15%) cases had recurrent vulvovaginal candidiasis (RVVC). The species distribution was as follows; Candida albicans (n = 21; 40.4%), C. glabrata (n = 11; 21.2%), C. tropicalis (n = 9; 17.3%), C. parapsilosis (n = 5; 9.7%), C. africana (n = 3; 5.7%), C. famata (n = 1; 1.9%), C. lusitaniae (n = 1; 1.9%), and C. dubliniensis (n = 1; 1.9%). The resistance rate of Candida isolates to fluconazole, itraconazole, and voriconazole were 15.38%, 11.5%, and 3.8%, respectively. Resistance to fluconazole was obtained in 46% (5/11) of RVVC cases but only in 7% (3/41) of VVC cases. CONCLUSION: This study demonstrated that the majority of VVC cases were caused by non-albicans Candida species which also were resistant to some antifungal agents. Hence, our findings revealed the importance of conducting periodical epidemiological studies to determine changes in species distribution. Moreover, for effective management of treatment and infection, it is imperative to evaluate the susceptibility profiles of Candida species isolated from VVC patients.
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Candidíase Vulvovaginal , Humanos , Feminino , Candidíase Vulvovaginal/tratamento farmacológico , Candidíase Vulvovaginal/epidemiologia , Candidíase Vulvovaginal/microbiologia , Antifúngicos/farmacologia , Irã (Geográfico)/epidemiologia , Fluconazol/farmacologia , Fluconazol/uso terapêutico , Estudos Transversais , Farmacorresistência Fúngica , Testes de Sensibilidade Microbiana , Candida albicansRESUMO
Leishmaniasis counts as one of the most neglected tropical diseases. Despite the amount of research perceived in this field, no fully effective and approved vaccine against this disease is yet available in humans. In this study, LACK and KMP11 antigens were constructed simultaneously by recombinant methods in prokaryotic and eukaryotic expression systems and were compared and assessed along with the CpG adjuvant in BALB/c mice. In the prokaryotic method, LACK and KMP11 protein gene sequences were synthesized in pET28a-TEV vector. In order to extract these two proteins after expression, His-tag and S-tag sequences were added to the constructs, respectively for LACK and KMP11. The pET28a-TEV-LACK/KMP11 construct was transformed into Escherichia coli, and the inserts were verified by Colony PCR. Pure proteins were verified by western blot, and groups of BALB/c mice were injected with the created prokaryotic recombinant proteins along with an ODN CpG adjuvant. In the eukaryotic method, antigen sequences were constructed in the pLEXSY-neo 2.1 vector, E.coli Top10 strain was cloned in the bacteria, and after being linearized were transfected into Leishmania tarentolae genome. After recombinant strains were selected, they were verified by molecular methods. After the extraction and purification of the protein using the method above, groups of mice were injected with the recombinant antigens and ODN CpG adjuvant. Eukaryotic subunit vaccines showed more effective immunization compared with prokaryotic vaccines and caused an immune system shift towards Th1 and protection. Protein expression in L. tarentolae by the constructs created in this host contains Post-Translational Modifications. The constructed protein will be significantly similar to eukaryotic proteins, considering that they are identical epitopes. More comprehensive studies aiming to improve the effectiveness of this vaccine are being conducted to improve immune profiles and immunological memory stimulation in future designs.
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Leishmania , Leishmaniose Cutânea , Animais , Eucariotos , Leishmania/genética , Leishmaniose Cutânea/prevenção & controle , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Recombinantes/genética , Vacinas de Subunidades Antigênicas/genéticaRESUMO
BACKGROUND: Candida species are known to cause serious fungal infections that produce cutaneous, mucosal, and systemic infections. Nowadays, mortality and morbidity candidiasis in immunocompromised patients have increased. Nanotechnology is a new world-known technology and includes particles ranging from about 1 to 100 nanometers. The purpose of this study was to evaluate the antifungal and cytotoxicity activities of titanium dioxide nanoparticles (TiO2-NPs) and zinc oxide nanoparticles (ZnO-NPs) compared to amphotericin B (AmB) on different Candida spp in in vitro conditions. METHODS: In the present study, susceptibility of different Candida species to TiO2-NPs and ZnO-NPs compared to AmB was determined by broth microdilution (BMD) and agar well diffusion methods. Cytotoxicity of TiO2-NPs and ZnO-NPs and amphotericin B was measured by MTT (3-(4, 5-Dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium Bromide) assay. RESULTS: The results indicated that the TiO2-NPs and ZnO-NPs showed antifungal activities against pathogenic Candida spp. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of TiO2-NP ranges against Candida spp. were 128-256 µg/mL and 256-512 µg/mL, respectively. The MIC and MFC values of ZnO-NPs were 64-128 µg/mL and 256-512 µg/mL, respectively. However, MICs and MFCs of AmB were 8-16 µg/mL and 16-32 µg/mL, respectively. The MTT assay results showed that the CC50% belonged to ZnO-NPs 706.2 µg/mL, for TiO2-NPs 862.1 µg/mL, and for AmB 70.19 µg/mL, respectively. CONCLUSION: Our findings showed that TiO2-NPs and ZnO-NPs had antifungal effects against all Candida species, yet the antifungal properties of TiO2-NPs and ZnO-NPs were significantly less than those of AmB. The CC50% of AmB was significantly lower than ZnO-NPs and TiO2-NPs.
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Anfotericina B , Antifúngicos , Candida/efeitos dos fármacos , Titânio , Óxido de Zinco , Anfotericina B/farmacologia , Anfotericina B/toxicidade , Animais , Antifúngicos/farmacologia , Antifúngicos/toxicidade , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Nanopartículas Metálicas/toxicidade , Camundongos , Testes de Sensibilidade Microbiana , Sais de Tetrazólio , Tiazóis , Titânio/farmacologia , Titânio/toxicidade , Óxido de Zinco/farmacologia , Óxido de Zinco/toxicidadeRESUMO
BACKGROUND: Drug resistance is a common phenomenon frequently observed in countries where leishmaniasis is endemic. Due to the production of the pteridine reductase enzyme (PTR1), drugs lose their efficacy, and consequently, the patient becomes unresponsive to treatment. This study aimed to compare the in vitro effect of meglumine antimoniate (MA) on non- healing Leishmania tropica isolates and on MA transfected non-healing one to PTR1. METHODS: Two non-healing and one healing isolates of L. tropica were collected from patients who received two courses or one cycle of intralesional MA along with biweekly liquid nitrogen cryotherapy or systemic treatment alone, respectively. After confirmation of L. tropica isolates by polymerase chain reaction (PCR), the recombinant plasmid pcDNA-rPTR (antisense) was transfected via electroporation and cultured on M199. Isolates in form of promastigotes were treated with different concentrations of MA and read using an enzyme-linked immunosorbent assay (ELISA) reader and the half inhibitory concentration (IC50 ) value was calculated. The amastigotes were grown in mouse macrophages and were similarly treated with various concentrations of MA. The culture glass slides were stained, and the mean number of intramacrophage amastigotes and infected macrophages were assessed in triplicate for both stages. RESULTS: All three transfected isolates displayed a reduction in optical density compared with the promastigotes in respective isolates, although there was no significant difference between non-healing and healing isolates. In contrast, in the clinical form (amastigotes), there was a significant difference between non-healing and healing isolates (p < 0.05). CONCLUSION: The results indicated that the PTR1 gene reduced the efficacy of the drug, and its inhibition by antisense and could improve the treatment of non-healing cases. These findings have future implications in the prophylactic and therapeutic modality of non- healing Leishmania isolates to drug.
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Leishmania tropica/genética , Leishmaniose Cutânea/tratamento farmacológico , Leishmaniose Cutânea/genética , Oxirredutases/genética , Proteínas de Protozoários/genética , Adulto , Animais , Antiprotozoários/farmacologia , Antiprotozoários/uso terapêutico , DNA Antissenso , Resistência Microbiana a Medicamentos/efeitos dos fármacos , Resistência Microbiana a Medicamentos/genética , Feminino , Humanos , Leishmania tropica/efeitos dos fármacos , Leishmania tropica/isolamento & purificação , Leishmaniose Cutânea/parasitologia , Masculino , Antimoniato de Meglumina/farmacologia , Antimoniato de Meglumina/uso terapêutico , Camundongos Endogâmicos BALB C , TransfecçãoRESUMO
BACKGROUND: Candida species are considered as the cause of one of the most important opportunistic fungal diseases. Accurate identification of Candida species is important because of antifungal susceptibility patterns are different among these species, so proper identification helps in the selection of antifungal drugs for the prevention and treatment. Phenotypic methods for identification of Candida species, which are widely used in clinical microbiology laboratories, have some limitations. Real-time PCR followed by the high-resolution melting analysis (HRMA) is a novel approach for the rapid recognition of pathogenic fungi. Molecular phylogeny is essential for obtaining a better understanding of the evolution of the genus Candida and the identification of the relative degree of the Candida species. The purpose of this study was molecular identification of Candida isolates by Real-time PCR-high-resolution melting analysis and investigation of the genetic diversity of Candida species. METHODS: Two hundred and thirty-two Candida isolates including 111 Candida isolates obtained from 96 HIV/AIDS patients and 121 Candida isolates obtained from 98 non-HIV persons were identified by real-time PCR and high-resolution melting curve analysis. To evaluate genetic diversity and relationships among Candida species, PCR products of nine clinical Candida isolates, as a representative of each kind of species, were randomly selected for DNA sequence analysis. RESULTS: In HIV/AIDS patients, six species of Candida spp. were identified as follows: C albicans (n = 64; 57.7%), C glabrata (n = 31; 27.92%), C parapsilosis (n = 9; 8.1%), C tropicalis (n = 4; 3.6%), C krusei (n = 2; 1.8%), and C kefyr (n = 1; 0.90%). In non-HIV persons, we identified eight species of Candida including C albicans (n = 46; 38.33%) followed by C glabrata and C krusei (each one, n = 18; 15%), C tropicalis (n = 13; 10.83%), C lusitaniae (n = 12; 5.17%), C parapsilosis (n = 10; 4.31%), and C kefyr and C guillermondii (each one, n = 2; 1.66%). Also, the phylogenetic analysis showed the presence of two main clades and six separate subclades. Accordingly, about 88.9% of the isolates were located in clade I and 11.10% of the studied isolates were in clade II. CONCLUSIONS: Real-time PCR followed by high-resolution melting analysis (HRMA) is known as a reliable, fast, and simple approach for detection and accurate identification of Candida species, especially in clinical samples.
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Candida/genética , Candida/isolamento & purificação , Variação Genética , Desnaturação de Ácido Nucleico/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Síndrome da Imunodeficiência Adquirida/microbiologia , Candida/classificação , DNA Fúngico/genética , Humanos , Filogenia , Padrões de Referência , Especificidade da EspécieRESUMO
BACKGROUND: Malaria is a public health concern that leads to about a million deaths worldwide every year. Malaria is caused by the genus Plasmodium, which includes P. falciparum, P. vivax, P. malariae, and P. ovale. Molecular phylogeny is essential to better recognition the evolution of the genus Plasmodium genus and detection of the relative degree of Plasmodium species in humans. The aim of this study was to detect malaria with Light Microscopy (LM) and Nested polymerase chain reaction (Nested PCR) methods in peripheral blood expansions and to investigate the genetic diversity of Plasmodium species by 18S rRNA gene in the southeast of Iran. METHODS: A total of 97 blood smears were collected from patients suspected to malaria in a 6-year period in the southeast of Iran including Hormozgan, Kerman, and Sistan and Baluchestan provinces. Diagnosis of Plasmodium species on blood smears was performed using LM and Nested PCR methods. In addition, 16 Plasmodium-positive samples were chosen for the determination of genetic diversity. RESULTS: Overall, 97 of 97 (100%) studied cases were positive by LM while 94 of 97 (96.8%) of them were detected as malaria by Nested PCR. Except for seven cases, Nested PCR confirmed the LM results. These samples involved two P. vivax and five P. falciparum in the LM method. Meanwhile, the Nested PCR was detected in all of the cases as a mixed infection with P. vivax and P. falciparum. The results of the phylogenetic analysis revealed two main clades and five different subclades. About 87.5% of the isolates were located in clade I and contained P. vivax. In addition, 12.5% of the studied isolates involved P. falciparum that was in clade II. CONCLUSION: According to our results, Nested PCR method had higher sensitivity than LM and is suggested as a good approach for malaria detection. Consideration the wide diversity of tested isolates and the importance of vaccine development, which is affected by this diversity, further studies are needed in this regard.
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DNA de Protozoário/genética , Malária/parasitologia , Microscopia/métodos , Plasmodium/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 18S/genética , Sangue/parasitologia , Feminino , Variação Genética , Humanos , Irã (Geográfico) , Malária/sangue , Masculino , Filogenia , Plasmodium/classificação , Plasmodium/citologia , Plasmodium/genéticaRESUMO
BACKGROUND: The present study was conducted to determine antimicotic susceptibility of Candida species (sp.) from patients with symptomatic candiduria. MATERIALS AND METHODS: Identification of Candida sp. and determination of efficacy of most routine antifungals were done using polymerase chain reaction-restriction fragment length polymorphism method and E-test, respectively. RESULTS: The results from susceptibility test reveal that caspofungin and amphotericin B have high antifungal activity against both albicans (100% and 96%, respectively) and nonalbicans (95.11% and 72.72%, respectively) isolates. CONCLUSION: The present study suggests that caspofungin and amphotericin B have the excellent ability to eradicate both Candida groups that showed decreased susceptibility to other compounds.
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OBJECTIVES: Vulvovaginal candidiasis (VVC) or vaginal candidiasis is a common fungal infection of the genitals causing inflammation, irritation, itching, and vaginal discharge. Common yeast infections are caused by the yeast species C. albicans. However, there are other species of Candida such as C. dubliniensis which are considered as the causative agents of this infection. Hydrolytic enzymes such as proteinase and coagulase are known as virulence factors. The aim of this study was the molecular confirmation and differentiation of C. dubliniensis among C. albicans strains isolated from women with vulvovaginal candidiasis by PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) and the evaluation of proteinase and coagulase activities. METHODS: A total of 100 C. albicans strains isolated from women with vulvovaginal candidiasis referred to Shiraz medical clinics were enrolled in the study. All the isolates were primarily identified by conventional methods. PCR-RFLP method was used for the confirmation and identification of C. albicans and C. dubliniensis. Moreover, in vitro proteinase and coagulase activities of these isolates were evaluated using bovine serum albumin media and classical rabbit plasma tube test. RESULTS: As a result, PCR-RFLP identified 100% of the isolates as C. albicans, and no C. dubliniensis could be identified in this study. 84% of the isolates showed proteinase activity, whereas coagulase activity was only detected in 5% of the isolates. CONCLUSIONS: This study reveals that C. dubliniensis plays no role in vaginal candidiasis in Iranian patients. Proteinase production could be an essential virulence factor in C. albicans pathogenicity, but coagulase activity has less potential in this matter.
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Candida albicans/enzimologia , Candida albicans/isolamento & purificação , Candida/enzimologia , Candida/isolamento & purificação , Candidíase Vulvovaginal/microbiologia , Candida/genética , Candida/patogenicidade , Candida albicans/genética , Candida albicans/patogenicidade , Candidíase Vulvovaginal/diagnóstico , Coagulase/análise , DNA Fúngico/isolamento & purificação , Feminino , Humanos , Irã (Geográfico) , Peptídeo Hidrolases/análise , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Fragmento de RestriçãoRESUMO
OBJECTIVES: Superficial and cutaneous fungal infections (SCFIs) are an important public health problem and are common in tropical and subtropical countries. Pityriasis versicolor, dermatophytosis, erythrasma, onychomycosis, and otomycosis are the major diseases associated with SCFIs. The aim of this study was to evaluate the prevalence and causative agents of dermatomycoses over a period of 10 years in Kerman province, southeast of Iran. METHODS: A number of 1782 subjects, including 1096 females and 686 males, with cutaneous disorders in their skin, nail, and hair suspected to have SCFIs participated in this study. The collected specimens were examined using direct microscopy examination, staining, culture on specific media and PCR-RFLP technique. RESULTS: In total, 617 (34.62%) subjects had SCFIs, of whom 290 (47%) were female and 327 (53%) were male. Identified SCFIs included yeast infections, dermatophytosis, saprophyte onychomycosis, erythrasma, and otomycosis due to non-dermatophytic molds (NDMs). The highest prevalence of dermatomycoses was found among the 41-50-year and 31-40-year age groups. Tinea unguium was the most common clinical pattern of dermatomycoses, and T. mentagrophytes was the predominant agent. Also, Aspergillus spp. were the most common NDMs agents of onychomycosis and otomycosis. CONCLUSIONS: This study summarized the epidemiological trends and etiologic agents of SCFIs in a 10-year period in Kerman, southeast of Iran. Consideration of the current epidemiologic trends in the prevalence and knowledge of the exact causative agents of SCFIs may play an important key role towards further investigations, diagnosis, and modification of current treatments.
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Dermatomicoses/epidemiologia , Dermatomicoses/microbiologia , Fungos/classificação , Fungos/isolamento & purificação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Irã (Geográfico)/epidemiologia , Masculino , Técnicas Microbiológicas , Microscopia , Pessoa de Meia-Idade , Técnicas de Diagnóstico Molecular , Prevalência , Estudos Retrospectivos , Adulto JovemRESUMO
Previously, we showed an inverse correlation between HSP27 serum levels and experimental atherogenesis in ApoE(-/-) mice that over-express HSP27 and speculated that the apparent binding of HSP27 to scavenger receptor-A (SR-A) was of mechanistic importance in attenuating foam cell formation. However, the nature and importance of the interplay between HSP27 and SR-A in atheroprotection remained unclear. Treatment of THP-1 macrophages with recombinant HSP27 (rHSP27) inhibited acLDL binding (-34%; p<0.005) and uptake (-38%, p<0.05). rHSP27 reduced SR-A mRNA (-39%, p=0.02), total protein (-56%, p=0.01) and cell surface (-53%, p<0.001) expression. The reduction in SR-A expression by rHSP27 was associated with a 4-fold increase in nuclear factor-kappa B (NF-κB) signaling (p<0.001 versus control), while an inhibitor of NF-κB signaling, BAY11-7082, attenuated the negative effects of rHSP27 on both SR-A expression and lipid uptake. To determine if SR-A is required for HSP27 mediated atheroprotection in vivo, ApoE(-/-) and ApoE(-/-) SR-A(-/-) mice fed with a high fat diet were treated for 3weeks with rHSP25. Compared to controls, rHSP25 therapy reduced aortic en face and aortic sinus atherosclerotic lesion size in ApoE(-/-) mice by 39% and 36% (p<0.05), respectively, but not in ApoE(-/-)SR-A(-/-) mice. In conclusion, rHSP27 diminishes SR-A expression, resulting in attenuated foam cell formation in vitro. Regulation of SR-A by HSP27 may involve the participation of NF-κB signaling. Lastly, SR-A is required for HSP27-mediated atheroprotection in vivo.
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Aorta/metabolismo , Aterosclerose/genética , Células Espumosas/metabolismo , Proteínas de Choque Térmico HSP27/genética , NF-kappa B/genética , Receptores Depuradores Classe A/genética , Animais , Aorta/efeitos dos fármacos , Aorta/patologia , Apolipoproteínas E/deficiência , Apolipoproteínas E/genética , Aterosclerose/metabolismo , Aterosclerose/patologia , Células CHO , Linhagem Celular , Cricetulus , Dieta Hiperlipídica , Modelos Animais de Doenças , Feminino , Células Espumosas/patologia , Regulação da Expressão Gênica , Proteínas de Choque Térmico HSP27/metabolismo , Proteínas de Choque Térmico/farmacologia , Humanos , Camundongos , Camundongos Knockout , Chaperonas Moleculares , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Proteínas de Neoplasias/farmacologia , Nitrilas/farmacologia , Receptores Depuradores Classe A/antagonistas & inibidores , Receptores Depuradores Classe A/metabolismo , Transdução de Sinais , Sulfonas/farmacologiaRESUMO
Background and Objectives: The aim of this study was to investigate the in vitro antifungal potency of the moronecidin-like peptide against Candida albicans, Candida glabrata, and Candida tropicalis. Materials and Methods: To evaluate the antifungal effect of moronecidin-like peptide, the protocol presented in CLSI M27-A3 and CLSI M27-S4 was used and the minimum inhibitory concentration was determined. Results: The minimum inhibitory effect of moronecidin-like peptide composition was 8 µg/ml for Candida tropicalis and Candida albicans and 32 µg/ml for Candida glabrata. The MIC of nystatin was determined to be 1.25 µg/ml for Candida glabrata and Candida albicans and 0.625 µg/ml for Candida tropicalis strains. The MFC composition of the moronecidin-like peptide was determined for Candida tropicalis and Candida albicans strains 8 µg/ml and for Candida glabrata strain 64 µg/ml. The results of cytotoxicity and hemolysis of the moronecidin peptide test on macrophage showed that moronecidin peptide has no cytotoxicity and toxicity properties. Conclusion: According to the results of the present study, the moronecidin-like peptide could be a new strategy in the treatment of infections caused by Candida strains. The discovery of the exact mechanism of which requires extensive clinical studies in this field.
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Pharmaceutical supply chain (PSC) is one of the most important healthcare supply chains and the recent pandemic (COVID-19) has completely proved it. Also, the environmental and social impacts of PSCs are undeniable due to the daily entrance of a large amount of pharmaceutical waste into the environment. However, studies on closed-loop PSCs (CLPSC) are rarely considered real-world requirements such as competition among diverse brands of manufacturers, the dependency of customers' demand on products' price and quality, and diverse reverse flows of end-of-life medicines. In this study, a scenario-based Multi-Objective Mixed-Integer Linear Programming model is developed to design a sustainable CLPSC, which investigates the reverse flows of expired medicines as three classes (must be disposed of, can be remanufactured and can be recycled). To study the competitive market and deal with demand uncertainty, a novel scenario-based game theory model is proposed. The demand function for each brand depends on the price and quality provided. Then, a hybrid solution approach is provided by combining the LP-metrics method with a heuristic algorithm. Furthermore, a real case study is investigated to evaluate the application of the model. Finally, sensitivity analysis and managerial insights are provided. The numerical results show that the proposed classification of reverse flows leads to proper waste management, making money, and reducing both disposal costs and raw material usage. Moreover, competition increases PSCs performance and improves the supply of products to pharmacies. Supplementary Information: The online version contains supplementary material available at 10.1007/s10479-021-03961-0.
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Background and Objectives: Candida albicans complex species are well known as the main cause of candidiasis, particularly among susceptible individuals. In this study, we report the genetic diversity of Candida spp. and the antifungal susceptibility pattern of the cryptic C. albicans complex isolates in Kerman, Iran. Materials and Methods: A total of 112 yeast isolates were obtained from different clinical samples, and molecular identification was performed. All C. albicans complex isolates were tested for susceptibility of them to amphotericin B, fluconazole, and itraconazole. Results: The majority of clinical isolates were C. albicans complex (n=48) followed by C. glabrata complex (n=34), C. parapsilosis complex (n=21), and C. krusei (n=9). Among C. albicans complex, 45 isolates were C. albicans (94%), 2 isolates were C. dubliniensis (4%), and 1 isolate was C. africana (2%). Amphotericin B was the most active antifungal, whereas 8.9% and 6.7% of the isolates were resistant to fluconazole and itraconazole, respectively. Conclusion: Regarding the high incidence of Candida infections particularly in susceptible populations and the emergence of an infrequent yeast species with elevated MICs, which is indistinguishable with conventional methods, developing accurate molecular methods for laboratory diagnosis should be considered in the clinical setting.
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Oropharyngeal Candidiasis (OPC) is an opportunistic fungal infection occurring in immunocompromised patients such as HIV/AIDS. The purpose of this study was to evaluate the antifungal properties of Lactobacillus acidophilus and Lactobacillus plantarum on different Candida species isolated from oral cavity of HIV/AIDS patients compared to Fluconazole (FLC). In this study, the antifungal effects of both cells and cell-free supernatants (CFSs) of L. acidophilus and L. plantarum were investigated against different oral Candida species by co-aggregation, agar overlay interference and broth microdilution assays, respectively. Our results showed that the highest co-aggregation ratio of the two tested Lactic acid bacteria (LAB) was observed for C. krusei. Both L. acidophilus and L. plantarum at cell concentrations 1010 to 102 cfu/ml were able to inhibit the growth of most of the oral Candida species, except for C. albicans, and to some C. krusei. In this study, MIC and MFC values for CFS of L. acidophilus ranged from 100 to 200 µl/ml and 100 to 200 µl/ml, respectively, and MIC and MFC values for CFS of L. plantarum were 50 to 200 µl/ml and 50 to 200 µl/ml, respectively. The ranges of MIC and MFC for FLC were 256-1024 µg/ml and 512-2048 µg/ml, respectively. C. albicans and C. parapsilosis displayed the highest and least susceptibility to CFSs of two LAB, respectively. Our findings showed that both cells and CFSs of L. acidophilus and L. plantarum had antifungal effects against oral Candida species.
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BACKGROUND: Leishmaniasis is a serious health problem in some parts of the world. In spite of the many known leishmaniasis control measures, the disease has continued to increase in endemic areas, and no effective vaccine has been discovered. METHODS: In this study, Leishmania tarentulae was used as a living factory for the production of two LACK and KMP11 immunogenic antigens in the mice body, and safety profiles were investigated. The sequences of the KMP11 and LACK L. major antigens were synthesized in the pLEXSY-neo 2.1 plasmid and cloned into E. coli strain Top10, and after being linearized with the SwaI enzyme, they were transfected into the genome of L. tarentolae. The L. tarentolae-LACK/KMP11/EGFP in the stationary phase with CpG ODN as an adjuvant was used for vaccination in BALB/c mice. Vaccination was performed into the left footpad. Three weeks later, the booster was injected in the same manner. To examine the effectiveness of the injected vaccine, pathogenic L. major (MRHO/IR/75/ER) was injected into the right footpad of all mice three weeks following the booster vaccination. In order to assess humoral immunity, the levels of IgG1, and IgG2a antibodies before and 6 weeks after the challenge were studied in the groups. In addition, in order to investigate cellular immunity in the groups, the study measured IFN-γ, IL-5, TNF-α, IL-6 and IL-17 cytokines before, 3 weeks and 8 weeks after the challenge, and also the parasite load in the lymph node with real-time PCR. RESULTS: The lowest level of the parasitic load was observed in the G1 group (mice vaccinated with L. tarentolae-LACK/KMP11/EGFP with CpG) in comparison with other groups (L. tarentolae-LACK/KMP11/EGFP +non-CpG (G2); L. tarentolae-EGFP + CpG (G3, control); L. tarentolae-EGFP + non-CpG (G4, control); and mice injected with PBS (G5, control). Moreover, the evaluation of immune response showed a delayed-type hypersensitivity towards Th1. CONCLUSIONS: According to the results of this study, the live recombinant vaccine of L. tarentolae-LACK/KMP11/EGFP with the CpG adjuvant reduced the parasitic load and footpad induration in infected mice. The long-term effects of this vaccine can be evaluated in volunteers as a clinical trial in future planning.
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Leishmania/imunologia , Vacinas contra Leishmaniose , Leishmaniose Cutânea , Vacinas Vivas não Atenuadas , Animais , Anticorpos Antiprotozoários , Antígenos de Protozoários/biossíntese , Antígenos de Protozoários/genética , Antígenos de Protozoários/imunologia , Clonagem Molecular , Citocinas/metabolismo , Escherichia coli/genética , Genes de Protozoários , Imunidade Humoral , Imunoglobulina G/metabolismo , Leishmania/efeitos dos fármacos , Leishmania/patogenicidade , Leishmania major/efeitos dos fármacos , Leishmania major/imunologia , Leishmania major/patogenicidade , Vacinas contra Leishmaniose/biossíntese , Vacinas contra Leishmaniose/imunologia , Vacinas contra Leishmaniose/farmacologia , Leishmaniose Cutânea/tratamento farmacológico , Leishmaniose Cutânea/imunologia , Leishmaniose Cutânea/prevenção & controle , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Camundongos Endogâmicos BALB C/parasitologia , Carga Parasitária , Proteínas de Protozoários/biossíntese , Proteínas de Protozoários/genética , Proteínas de Protozoários/imunologia , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/imunologia , Vacinas Vivas não Atenuadas/biossíntese , Vacinas Vivas não Atenuadas/imunologia , Vacinas Vivas não Atenuadas/farmacologia , Vacinas Sintéticas/biossíntese , Vacinas Sintéticas/imunologia , Vacinas Sintéticas/farmacologiaRESUMO
Estradiol and related estrogens have been widely used as supplements to relieve menopausal symptoms, but they lead to an increased risk of breast and endometrial cancer. Here we report the synthesis of a new family of compounds where we have removed the B-ring from the steroid ABCD structure, and functionalized the A-ring. These A-CD compounds show a preferential affinity for the estrogen receptor subtype ERbeta. Some show binding affinities which are greater than estradiol. The presence of electron-withdrawing substituents on the A-ring should reduce the tendency of these compounds to form carcinogenic metabolites, so they might lead to a safer approach to hormone replacement therapy.
Assuntos
Estradiol , Receptores de Estradiol/agonistas , Estradiol/agonistas , Estradiol/análogos & derivados , Estradiol/síntese química , Estradiol/farmacologia , Feminino , Terapia de Reposição Hormonal/métodos , Humanos , Modelos Moleculares , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
BACKGROUND AND PURPOSE: Candida species are the common opportunistic pathogens during the course of human immunodeficiency virus (HIV) infection. Oropharyngeal candidiasis (OPC) is generally known as the initial sign of HIV infection. The aim of this study was to compare demographic characteristics and frequency of Candida species between HIV/AIDS patients and non-HIV subjects in Kerman, southeast of Iran. MATERIALS AND METHODS: This study was conducted on 143 samples collected from the oral cavity of 81 HIV/AIDS patients and 35 non-HIV subjects. The samples were cultured on Sabouraud dextrose agar and CHROMagar. The identification of Candida species was accomplished using the color of colony and polymerase chain reaction-restriction fragment length polymorphism. RESULTS: According to the results, C. albicans (n=25, 69.14%) was the most prevalent species isolated from the HIV/AIDS patients, followed by C. glabrata (n=19, 23.46%). Other isolated species included C. parapsilosis (n=4, 4.94 %), C. krusei (n=1, 1.24%), and C. kefyr (n=1, 1.24%). Out of the 35 Candida species recovered from the oral samples of non-HIV subjects, 23 (65.71%) and 12 (34.29%) cases were C. krusei and C. albicans, respectively. Candida krusei was the only non-albicans species found in the non-HIV subjects that was also the predominant isolated species. Regarding the HIV/AIDS patients, the highest prevalence of OPC was observed in the age group of 41-50 years. However, in the non-HIV subjects, the age group of 31-40 years had the highest prevalence of this infection. Furthermore, no correlation was observed between the gender and number of Candida isolates. CONCLUSION: Consideration of the epidemiologic data showed that the two groups were significantly different in terms of the prevalence of Candida species, which could play a major role in the selection of effective drugs for the treatment of candidiasis.
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Phenol is a hazardous organic chemical that introduced into the environment by industrial and pharmaceutical discharges. As a versatile option for phenol removal, adsorption would be viable if it accompanying with low cost adsorbents. This article described a natural, very cheap and local available adsorbent for phenol removal. Phenol showed a high affinity to Citrullus colocynthis waste ash which mainly composed of SiO2 (41.6%), Al2O3 (17.3%) and MgO (15.9%). Up to 70% of phenol adsorbed in the first 30â¯min of agitation. The phenol removal was increased by increasing adsorbent dose (0.5-10â¯g/L) and decreasing pH (2-12) and pollutant concentration (10-100â¯mg/L). The positive value of ∆H° in thermodynamic data (0.06) revealed that the process is endothermic. The high and positive value of ∆S° (13.01) and negative values of ∆G° (- 5.36 to - 7.28), showed a high affinity of phenol to the adsorbent and the spontaneous nature of the adsorption. Isotherm modelling revealed that the phenol molecules adsorbed in multilayer with the maximum adsorption capacity of 173.2â¯mg/g. The rate limiting step in the sorption process was chemisorption, based on the kinetic data.